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Analytical Chem JoVE (Open Access) Citable · DOI

Resurrection of Dormant Daphnia magna: Protocol and Applications

DOI: 10.3791/56637-v
What you'll learn
  • ✓ Prepare and radiometrically date sediment cores for temporal analysis
  • ✓ Extract and decapsulate Daphnia ephippia from sediment layers
  • ✓ Resurrect dormant Daphnia magna and establish isoclonal laboratory lines
  • ✓ Generate longitudinal evolutionary data from sedimentary archives
Protocol

Long-term studies are essential to understanding the process of evolution and the mechanisms of adaptation. Generally, these studies require commitments beyond the life-time of researchers. Here, a powerful method is described that dramatically advances state-of-the-art data collection to generate longitudinal data in natural systems.

Difficulty
advanced
Total time
~2–4 weeks (including sediment preparation, radiometric dating, hatching, and isoclonal line establishment)
Model organism
Daphnia magna
Biosafety
BSL-1

Steps

1
Prepare and radiometrically date sediment cores

Obtain sediment cores from natural water bodies and perform radiometric dating to establish chronology. This establishes the temporal framework for subsequent fossil resurrection experiments.

▶ 01:08
2
Sieve sediment layers into size fractions

Separate sediment core material into discrete size fractions using mesh sieves. This concentrates ephippia and removes bulk sediment before hatching.

▶ 02:01
3
Decapsulate ephippia and hatch Daphnia

Remove dormant eggs (ephippia) from sediment, decapsulate to expose resting embryos, and induce hatching under controlled laboratory conditions. Establish isoclonal lines from individual hatched organisms.

▶ 03:23
4
Assess resurrected Daphnia phenotypic responses

Culture resurrected subpopulations under standardized conditions and measure phenotypic traits (e.g., temperature tolerance) to characterize evolutionary changes across time.

▶ 05:16
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